Anti-GCLM antibody [EPR6667]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (AB126704)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder cancer tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IP

Unknown

Immunoprecipitation - Anti-GCLM antibody [EPR6667] (AB126704)

Purified ab126704 at 1/20 dilution (0.6μg) immunoprecipitating GCLM in HeLa whole cell lysate.
Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+) : ab126704 + HeLa whole cell lysate.
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab126704 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
Blocking Buffer and concentration : 5% NFDM/TBST.
Diluting buffer and concentration : 5% NFDM/TBST.
Observed band size : 31 kDa

All lanes:

Immunoprecipitation - Anti-GCLM antibody [EPR6667] (ab126704)

Predicted band size: 31 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (AB126704)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Rat liver tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-GCLM antibody [EPR6667] (AB126704)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Mouse cardiac muscle tissue sections labeling GCLM with purified ab126704 at 1/50 dilution (2.4 µg/mL). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• WB

Lab

Western blot - Anti-GCLM antibody [EPR6667] (AB126704)

Lanes 1 - 3 : Merged signal (red and green). Green - ab126704 observed at 31 kDa. Red - loading control, ab130007, observed at 130 kDa.

ab126704 was shown to recognize GCLM in wild-type HAP1 cells as signal was lost at the expected MW in GCLM knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and GCLM knockout samples were subjected to SDS-PAGE. ab126704 and ab130007 (Mouse anti-Vinculin loading control) were incubated overnight at 4°C at 1 μg/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-GCLM antibody [EPR6667] (ab126704) at 1 µg/mL

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

GCLM knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa whole cell lysate at 20 µg

Predicted band size: 31 kDa

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• WB

Unknown

Western blot - Anti-GCLM antibody [EPR6667] (AB126704)

All lanes:

Western blot - Anti-GCLM antibody [EPR6667] (ab126704) at 1/10000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma ) whole cell lysate

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 31 kDa

false